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K Leary

Publications and source records attributed to K Leary.

15 recordsLinked to original sources
PubMed1997

Race, self-disclosure, and "forbidden talk": race and ethnicity in contemporary clinical practice.

In this paper I attempt to extend the psychoanalytic conversation about race and ethnicity by discussing the intersubjectivity of race and racial difference. I present clinical material from an interracial treatment in which disclosures about race played an important role in deepening the clinical process. The resulting interactions permitted the patient to admit more of herself into the treatment space. I suggest that contemporary psychoanalytic formulations and multicultural perspectives from outside of psychoanalysis can together create more meaningful conceptualizations which take into account the lived realities of race and the ways in which these may be shaped by individual psychology.

Adult↗
PubMed1994

Psychoanalytic "problems" and postmodern "solutions".

A number of recent revisions of psychoanalytic theory implicitly draw on postmodern conceptualizations of human selves and human subjectivity. Though postmodern ideas have a wide currency in the humanities and in literary criticism, and are increasingly represented in critiques of science, psychoanalytic clinicians are generally less familiar with the body of writings that encompass postmodernist thought. This paper discusses the evolution of postmodernism and its emergence into psychoanalytic theory using the work of Roy Schafer and Irwin Hoffman as cases in point. I will suggest that when postmodernism is applied to psychoanalytic practice, the result is only a partial solution, at best, to the problems of metapsychology postmodernist revisions were intended to resolve.

Culture↗
PubMed1993

Regulation of herpes simplex virus type 1 gene expression in nonpermissive murine resident peritoneal macrophages.

The restriction of herpes simplex virus type 1 (HSV-1) gene expression in nonpermissive murine resident peritoneal macrophages (ResPMO) was analyzed at the level of transcription. Using nuclear run-on assays, we established that transcription of a representative gene from each HSV-1 kinetic class was initiated in both ResPMO and permissive Vero cells. Using transient expression assays, we observed that the promoters of two early genes were activated by the immediate-early protein ICP4 in ResPMO. However, the level of transactivation of these promoters by another immediate-early protein, ICP0, was markedly reduced in ResPMO. Furthermore, the synergistic trans-activation normally observed between ICPs 4 and 0 in Vero cells was noticeably absent in ResPMO. The data indicate that the block in viral gene expression in ResPMO occurs after initiation of viral gene transcription and involves dysfunction of a viral immediate-early regulatory protein, ICP0.

Animals↗
PubMed1992

Analysis of the basis for persistence of herpes simplex virus type 1 in undifferentiated U937 cells.

Replication of herpes simplex type 1 (HSV-1) is inhibited in the human monocyte-like cell line, U937, when the cells are in the undifferentiated state, but when the cells are stimulated to differentiate by treatment with the phorbol ester, phorbol 12-myristate 13-acetate virus is replicated. Because HSV-1 has been shown to persist in these cells and in their in vitro counterparts freshly isolated human blood monocytes, we initiated an analysis of viral persistence in undifferentiated U937 cells. No appreciable HSV-1 DNA replication was observed in undifferentiated U937 cells compared with differentiated U937 cells and with fully permissive Vero cells. However, using in situ hybridization, we established that a significant percent of the undifferentiated U937 cells contained viral DNA sequences. Interestingly, when analyzed by Southern blot hybridization, this DNA was found to have assumed a nonlinear configuration similar to that found in latently infected neurons. Analysis of viral proteins in undifferentiated U937 cells revealed a marked absence of proteins of all three kinetic classes. However, in transient transfection assays, the major viral transactivating protein ICP4, functioned normally, whereas ICP0, a promiscuous transactivator of both viral and cellular genes, was unable to transactivate viral promoters in undifferentiated U937 cells. Thus, a subtle dysfunction in the activity of ICP0 may account, at least in part, for the inability of undifferentiated U937 cells to support replication of HSV-1.

Animals↗
PubMed1990

Transient expression of virus-specific promoters in murine resident peritoneal macrophages.

Monocyte-macrophages (MO), being non-permissive for most viruses, play an important role in resistance to virus infection. In order to establish the mechanism of abortive infection of murine resident peritoneal MO (ResPMO) by herpes simplex virus type 1 (HSV-1), it is desirable to transfect these cells with viral promoters linked to an assayable gene, for example, the bacterial chloramphenicol acetyl transferase (CAT) gene. This will facilitate studies designed to measure levels of promoter activation or repression in these specialized cells. Transient expression of CAT in ResPMO was achieved with DEAE-dextran, but not using either calcium phosphate precipitate or lipofectin. CAT expression driven by various virus-specific promoters was less efficient in ResPMO compared with Vero cells and approximately 50% of input plasmid DNA remained in Vero cells at 48 h post transfection, but only 9% was detectable in ResPMO. However, approximately 6% of ResPMO and 9% of Vero cells contained CAT-specific DNA at 24 h post transfection. In addition, 2% of cells of either cell type contained CAT-specific polypeptide at 48 h. This is therefore the first report that the non-replicating murine ResPMO can be transfected in vitro and more importantly, that these cells express the transfected gene products.

Animals↗
PubMed1989

The influence of the herpes simplex virus-1 DNA template environment on the regulation of gene expression.

To determine the role of the HSV-1 genome structure and environment on the regulation of gene expression, we constructed recombinant viruses containing a heterologous gene inserted into either the immediate early ICPO or late glycoprotein C (gC) genes of HSV-1. The heterologous gene consisted of the SV40 early promoter (without enhancer sequences) linked to the coding sequences for the bacterial chloramphenicol acetyl transferase (CAT). The expression of CAT was examined in Vero cells infected with either virus (named ICP0-CAT and Sph 6). For both recombinants, expression of CAT was not dependent upon prior viral protein synthesis. The kinetics of expression of CAT-specific mRNA resembled that of the HSV-1 genes into which CAT was inserted. Primer extension analysis revealed that the SV40 promoter is recognized and used when placed in cis in two different HSV-1 genome locations, and Northern hybridization experiments confirmed that the heterologous gene was expressed in the absence of prior viral protein synthesis. Therefore, this gene was not regulated as strictly as an HSV-1 gene, but was influenced by the environment into which it was placed, presumably by factors that are present when the normal viral gene is on.

Base Sequence↗
PubMed1989

Molecular localization of abortive infection of resident peritoneal macrophages by herpes simplex virus type 1.

Mononuclear phagocytes exhibit different patterns of intrinsic resistance to herpes simplex virus type 1 (HSV-1) that are related to the heterogeneity of macrophage populations and may reflect the particular differentiation or maturation state of the macrophages. In this study, we characterized the molecular basis for the block in HSV-1 replication in resident peritoneal macrophages from B6C3F1 mice. Infected resident peritoneal macrophages were analyzed for the presence of virus-specific mRNA by Northern (RNA) blotting and in situ hybridization and for proteins by immunofluorescence. The data were compared with those obtained in HSV-1-infected permissive Vero cells. The immediate-early genes ICP4, ICP0, ICP22, and ICP27 were transcribed in resident peritoneal macrophages, as was the early gene tk. Virus-specific mRNA for the major DNA-binding protein ICP8 was barely detectable, and that for another early gene, the viral DNA polymerase, was not detected. In addition, transcripts for the delayed-early gene glycoprotein D and the true late gene glycoprotein C (gC) were not detectable in resident peritoneal macrophages. In situ hybridization and immunofluorescence studies confirmed that transcripts and proteins for the immediate-early and some early HSV-1 genes were present. These data also established that 14% of the resident peritoneal macrophages were positive for RNA and polypeptide specific for the immediate-early gene ICP4 and that 7 to 11% were positive for RNA or polypeptides specific for the early genes tk and ICP8. The fact that only a few cells expressed viral products emphasizes the heterogeneity that exists even in this relatively homogeneous resident peritoneal macrophage population. Consistent with the Northern blot analysis, no RNA specific for the early DNA polymerase gene or the late gC gene was detected by in situ hybridization nor could the polypeptide for the gC gene be seen by immunofluorescence. Thus, while early transcriptional events were initiated in some resident peritoneal macrophages, there was a block in replication localized at the level of expression of the early to delayed-early viral genes.

Animals↗
PubMed1988

The herpes simplex virus type 1 alpha protein ICP27 can act as a trans-repressor or a trans-activator in combination with ICP4 and ICP0.

The herpes simplex virus type 1 (HSV-1) alpha proteins ICP4, ICP0, and ICP27 are trans-acting proteins which affect HSV-1 gene expression. To investigate potential interactions between these alpha products and to determine the specificity of action of the alpha proteins in combination with each other compared with their activities individually, we performed a series of transient-expression assays. In these assays we used plasmids containing the alpha genes encoding ICP4, ICP0, and ICP27 either singly or in combination as effectors and HSV-1 genes of different kinetic classes and heterologous genes as targets. The HSV-1 targets consisted of promoter-regulatory domains from alpha (ICP0 and ICP27), beta (thymidine kinase and alkaline exonuclease), beta-gamma (glycoprotein D, glycoprotein B, and VP5), and gamma (glycoprotein C) genes, each fused to the chloramphenicol acetyltransferase (CAT) gene. The heterologous target genes consisted of the simian virus 40 early promoter with enhancer and the Rous sarcoma virus long terminal repeat promoter and enhancer each fused to the CAT gene. Target promoter activity was measured by the assay of CAT activity in extracts of transfected cells and by Northern (RNA) blot hybridization of CAT mRNA. The results of these experiments showed that ICP4 activated only HSV-1 target genes, whereas ICP0 activated all of the targets and ICP27 had little effect on any of the targets. ICP4 and ICP0 had a synergistic effect when inducing HSV-1 targets, but they did not have this effect on the heterologous targets pSV2-CAT or pRSV-CAT. In fact, lower levels of CAT activity and CAT mRNA were found in the presence of both effectors than with ICP0 alone. Most interestingly, although the effector plasmid containing the ICP27 gene had little effect on its own, two different and marked effects depending on the target were observed when ICP27 was combined with ICP4 or ICP0 or both. A trans-repression of the induction seen with ICP4 and ICP0 was found when ICP27 was present in the transfections with pSV2-CAT, pRSV-CAT, pICP0-CAT, pICP27-CAT, pTK-CAT, pgD-CAT, pgB-CAT, and pgC-CAT. This resulted in CAT activity levels which were similar to or lower than the basal level of expression of the target genes in the absence of effector plasmids. This trans-repression occurred over a wide range of concentrations of input ICP27 plasmid. In contrast to this repressive effect of ICP27, a trans-activation was seen when ICP4, ICP0, and ICP27 plasmids were combined in transfections with pAE-CAT and pVP5-CAT as targets. This trans-activation also occurred over a 10-fold range of input ICP27 plasmid. These results suggest that ICP27 can facilitate both down

Animals↗
PubMed1987

The alpha protein ICP0 does not appear to play a major role in the regulation of herpes simplex virus gene expression during infection in tissue culture.

The herpes simplex virus type 1 (HSV-1) alpha protein ICP0 trans-activates HSV-1 early genes in transient expression assays. To investigate the function of ICP0 during HSV-1 infection, we have lowered the level of ICP0 by use of a recombinant plasmid that has been engineered to express the antisense message. Cell lines were constructed which stably carry the antisense plasmid. Total protein profiles from infected antisense cell lines showed that the level of ICP0 was reduced to less than 10% of the wild type level in two of the cell lines. However, reducing the level of ICP0 did not have a significant effect on the expression of HSV-1 early or late genes. The polypeptide patterns for the remaining infected cell polypeptides were similar in that no bands were absent although there were some quantitative differences. The level of two early proteins, glycoprotein B and glycoprotein D was reduced in one of the cell lines, however, levels were nearly equivalent to the control infection for two other cell lines tested. Virus yields were the same for the antisense cell lines and for parent cells. Decreased ICP0 levels did not lead to more restrictive phenotypes for an alpha 4 or alpha 27 mutant as protein patterns were similar for these mutants in antisense and parent cells. Therefore, while ICP0 has been demonstrated to be a strong inducer of gene expression in transient expression assays, it does not appear to have a major role as an activator during the productive infection of tissue culture cells.

Animals↗
PubMed1985

Comparison of herpes simplex virus type 1 DNA replication and virus production in murine bone marrow-derived and resident peritoneal macrophages.

The mechanism of resistance of murine macrophages (M phi) to infection by herpes simplex virus type 1 (HSV-1) was examined. Infection of bone marrow-derived M phi (BMDM phi) and resident peritoneal M phi (Res-M phi) was compared with infection of permissive Vero cells. In contrast to HSV-1 infection in Vero cells, no infectious virus was produced from either M phi cell type. However, marked cytopathic effect (c.p.e.) was evident in BMDM phi at 48 h post-infection, while there was no c.p.e. at any time post-infection in the Res-M phi. Cloned EcoRI subgenomic fragments representing the entire HSV-1 genome were used as probes in DNA :DNA hybridization experiments to determine the viral genome content in the infected cell types. In Res-M phi, HSV-1 DNA was present at early times post-infection but declined rapidly. In BMDM phi, the virus genome was always detected and increased with time after infection. The results suggest that Res-M phi restrict HSV-1 production at a point prior to viral DNA synthesis, whereas the block in HSV production in BMDM phi occurs at a later stage in the viral replicative cycle.

Animals↗
PubMed1984

The interaction of a topoisomerase-like enzyme from herpes simplex virus type 1-infected cells with non-viral circular DNA.

An enzyme activity from herpes simplex virus type 1 (HSV-1)-infected baby hamster kidney cells has been identified which generates large networks of pBR322 DNA from a monomeric DNA substrate. Extracts derived from cells infected at the non-permissive temperature, with the early regulatory mutants of HSV-1, tsK and tsB2, did not contain activity, suggesting that the enzyme is virus-induced and may be virus-specific. The enzyme is similar to the DNA topoisomerases in that network formation was dependent upon the presence of Mg2+ and a DNA condensing agent, and ATP was not required. Following digestion with EcoRI, the networks could be resolved to a single, linear, monomeric species of pBR322 DNA.

Animals↗
PubMed1983

Replication of herpes simplex virus type 1 on hydroxyurea-resistant baby hamster kidney cells.

Hydroxyurea-resistant (HUr) baby hamster kidney cells were isolated, subcloned, and characterized. One clonal line, which contained elevated levels of ribonucleotide reductase, lost its HU resistance during passage in the absence of the inhibitor, whereas another clonal line was stably resistant. The replication of herpes simplex virus type 1 on these cells was compared with that of the parvovirus minute virus of mice. Herpes simplex virus type 1 was found to be as sensitive to HU on both lines of HUr baby hamster kidney cells as it was on parental (HU-sensitive) cells, whereas parvovirus replication was about eight times more resistant on HUr baby hamster kidney cells compared with the parental cells. The results suggest that herpes simplex virus type 1 cannot use the cellular reductase and may code for its own.

Animals↗